Metastasis is responsible for >90% of cancer-related deaths. Complex signaling in cancer cells orchestrates the progression from a primary to a metastatic cancer. However, the mechanisms of these cellular changes remain elusive. We previously demonstrated that p90 ribosomal S6 kinase 2 (RSK2) promotes tumor metastasis. Here we investigated the role of RSK2 in the regulation of microtubule dynamics and its potential implication in cancer cell invasion and tumor metastasis. Stable knockdown of RSK2 disrupted microtubule stability and decreased phosphorylation of stathmin, a microtubule-destabilizing protein, at serine 16 in metastatic human cancer cells. We found that RSK2 directly binds and phosphorylates stathmin at the leading edge of cancer cells. Phosphorylation of stathmin by RSK2 reduced stathmin-mediated microtubule depolymerization. Moreover, overexpression of phospho-mimetic mutant stathmin S16D significantly rescued the decreased invasive and metastatic potential mediated by RSK2 knockdown in vitro and in vivo. Furthermore, stathmin phosphorylation positively correlated with RSK2 expression and metastatic cancer progression in primary patient tumor samples. Our finding demonstrates that RSK2 directly phosphorylates stathmin and regulates microtubule polymerization to provide a pro-invasive and pro-metastatic advantage to cancer cells. Therefore, the RSK2–stathmin pathway represents a promising therapeutic target and a prognostic marker for metastatic human cancers.
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Fidler IJ . The pathogenesis of cancer metastasis: the 'seed and soil' hypothesis revisited. Nat Rev Cancer 2003; 3: 453–458.
Nguyen DX, Massague J . Genetic determinants of cancer metastasis. Nat Rev Genet 2007; 8: 341–352.
Gupta GP, Massague J . Cancer metastasis: building a framework. Cell 2006; 127: 679–695.
Sahai E . Mechanisms of cancer cell invasion. Curr Opin Genet Dev 2005; 15: 87–96.
Li Y, Wang JP, Santen RJ, Kim TH, Park H, Fan P et al. Estrogen stimulation of cell migration involves multiple signaling pathway interactions. Endocrinology 2010; 151: 5146–5156.
Persad S, Dedhar S . The role of integrin-linked kinase (ILK) in cancer progression. Cancer Metastasis Rev 2003; 22: 375–384.
Jin L, Li D, Lee JS, Elf S, Alesi GN, Fan J et al. p90 RSK2 mediates antianoikis signals by both transcription-dependent and -independent mechanisms. Mol Cell Biol 2013; 33: 2574–2585.
Kang S, Elf S, Lythgoe K, Hitosugi T, Taunton J, Zhou W et al. p90 ribosomal S6 kinase 2 promotes invasion and metastasis of human head and neck squamous cell carcinoma cells. J Clin Invest 2010; 120: 1165–1177.
Li D, Jin L, Alesi GN, Kim YM, Fan J, Seo JH et al. The prometastatic ribosomal S6 kinase 2-cAMP response element-binding protein (RSK2-CREB) signaling pathway up-regulates the actin-binding protein fascin-1 to promote tumor metastasis. J Biol Chem 2013; 288: 32528–32538.
Buck M, Poli V, Hunter T, Chojkier M . C/EBPbeta phosphorylation by RSK creates a functional XEXD caspase inhibitory box critical for cell survival. Mol Cell 2001; 8: 807–816.
Palmer A, Gavin AC, Nebreda AR . A link between MAP kinase and p34(cdc2)/cyclin B during oocyte maturation: p90(rsk) phosphorylates and inactivates the p34(cdc2) inhibitory kinase Myt1. EMBO J 1998; 17: 5037–5047.
Shimamura A, Ballif BA, Richards SA, Blenis J . Rsk1 mediates a MEK-MAP kinase cell survival signal. Curr Biol 2000; 10: 127–135.
Dehan E, Bassermann F, Guardavaccaro D, Vasiliver-Shamis G, Cohen M, Lowes KN et al. betaTrCP- and Rsk1/2-mediated degradation of BimEL inhibits apoptosis. Mol Cell 2009; 33: 109–116.
Blenis J . Signal transduction via the MAP kinases: proceed at your own RSK. Proc Natl Acad Sci USA 1993; 90: 5889–5892.
Frodin M, Gammeltoft S . Role and regulation of 90 kDa ribosomal S6 kinase (RSK) in signal transduction. Mol Cell Endocrinol 1999; 151: 65–77.
Anjum R, Blenis J . The RSK family of kinases: emerging roles in cellular signalling. Nat Rev Mol Cell Biol 2008; 9: 747–758.
Schoumacher M, Goldman RD, Louvard D, Vignjevic DM . Actin, microtubules, and vimentin intermediate filaments cooperate for elongation of invadopodia. J Cell Biol 2010; 189: 541–556.
Desai A, Mitchison TJ . Microtubule polymerization dynamics. Annu Rev Cell Dev Biol 1997; 13: 83–117.
Belmont LD, Mitchison TJ . Identification of a protein that interacts with tubulin dimers and increases the catastrophe rate of microtubules. Cell 1996; 84: 623–631.
Belletti B, Nicoloso MS, Schiappacassi M, Berton S, Lovat F, Wolf K et al. Stathmin activity influences sarcoma cell shape, motility, and metastatic potential. Mol Biol Cell 2008; 19: 2003–2013.
Tan HT, Wu W, Ng YZ, Zhang X, Yan B, Ong CW et al. Proteomic analysis of colorectal cancer metastasis: stathmin-1 revealed as a player in cancer cell migration and prognostic marker. J Proteome Res 2012; 11: 1433–1445.
Hsieh SY, Huang SF, Yu MC, Yeh TS, Chen TC, Lin YJ et al. Stathmin1 overexpression associated with polyploidy, tumor-cell invasion, early recurrence, and poor prognosis in human hepatoma. Mol Carcinog 2010; 49: 476–487.
Cheng AL, Huang WG, Chen ZC, Peng F, Zhang PF, Li MY et al. Identification of novel nasopharyngeal carcinoma biomarkers by laser capture microdissection and proteomic analysis. Clin Cancer Res 2008; 14: 435–445.
Jeon TY, Han ME, Lee YW, Lee YS, Kim GH, Song GA et al. Overexpression of stathmin1 in the diffuse type of gastric cancer and its roles in proliferation and migration of gastric cancer cells. Br J Cancer 2010; 102: 710–718.
Polzin RG, Benlhabib H, Trepel J, Herrera JE . E2F sites in the Op18 promoter are required for high level of expression in the human prostate carcinoma cell line PC-3-M. Gene 2004; 341: 209–218.
Kinoshita I, Leaner V, Katabami M, Manzano RG, Dent P, Sabichi A et al. Identification of cJun-responsive genes in Rat-1a cells using multiple techniques: increased expression of stathmin is necessary for cJun-mediated anchorage-independent growth. Oncogene 2003; 22: 2710–2722.
Carr JR, Park HJ, Wang Z, Kiefer MM, Raychaudhuri P . FoxM1 mediates resistance to herceptin and paclitaxel. Cancer Res 2010; 70: 5054–5063.
San-Marina S, Han Y, Liu J, Minden MD . Suspected leukemia oncoproteins CREB1 and LYL1 regulate Op18/STMN1 expression. Biochim Biophys Acta 2012; 1819: 1164–1172.
Li N, Jiang P, Du W, Wu Z, Li C, Qiao M et al. Siva1 suppresses epithelial-mesenchymal transition and metastasis of tumor cells by inhibiting stathmin and stabilizing microtubules. Proc Natl Acad Sci USA 2011; 108: 12851–12856.
Daub H, Gevaert K, Vandekerckhove J, Sobel A, Hall A . Rac/Cdc42 and p65PAK regulate the microtubule-destabilizing protein stathmin through phosphorylation at serine 16. J Biol Chem 2001; 276: 1677–1680.
Marklund U, Larsson N, Brattsand G, Osterman O, Chatila TA, Gullberg M . Serine 16 of oncoprotein 18 is a major cytosolic target for the Ca2+/calmodulin-dependent kinase-Gr. Eur J Biochem 1994; 225: 53–60.
Beretta L, Dobransky T, Sobel A . Multiple phosphorylation of stathmin. Identification of four sites phosphorylated in intact cells and in vitro by cyclic AMP-dependent protein kinase and p34cdc2. J Biol Chem 1993; 268: 20076–20084.
Cassimeris L . The oncoprotein 18/stathmin family of microtubule destabilizers. Curr Opin Cell Biol 2002; 14: 18–24.
Belletti B, Baldassarre G . Stathmin: a protein with many tasks. New biomarker and potential target in cancer. Expert Opin Ther Targets 2011; 15: 1249–1266.
Misek DE, Chang CL, Kuick R, Hinderer R, Giordano TJ, Beer DG et al. Transforming properties of a Q18—>E mutation of the microtubule regulator Op18. Cancer Cell 2002; 2: 217–228.
Holmfeldt P, Brannstrom K, Stenmark S, Gullberg M . Aneugenic activity of Op18/stathmin is potentiated by the somatic Q18—>e mutation in leukemic cells. Mol Biol Cell 2006; 17: 2921–2930.
Kang Y, He W, Tulley S, Gupta GP, Serganova I, Chen CR et al. Breast cancer bone metastasis mediated by the Smad tumor suppressor pathway. Proc Natl Acad Sci USA 2005; 102: 13909–13914.
Tournebize R, Andersen SS, Verde F, Doree M, Karsenti E, Hyman AA . Distinct roles of PP1 and PP2A-like phosphatases in control of microtubule dynamics during mitosis. EMBO J 1997; 16: 5537–5549.
Mistry SJ, Li HC, Atweh GF . Role for protein phosphatases in the cell-cycle-regulated phosphorylation of stathmin. Biochem J 1998; 334: 23–29.
Mizumura K, Takeda K, Hashimoto S, Horie T, Ichijo H . Identification of Op18/stathmin as a potential target of ASK1-p38 MAP kinase cascade. J Cell Physiol 2006; 206: 363–370.
Kang S, Dong S, Gu TL, Guo A, Cohen MS, Lonial S et al. FGFR3 activates RSK2 to mediate hematopoietic transformation through tyrosine phosphorylation of RSK2 and activation of the MEK/ERK pathway. Cancer Cell 2007; 12: 201–214.
Clark DE, Poteet-Smith CE, Smith JA, Lannigan DA . Rsk2 allosterically activates estrogen receptor alpha by docking to the hormone-binding domain. EMBO J 2001; 20: 3484–3494.
Hitosugi T, Fan J, Chung TW, Lythgoe K, Wang X, Xie J et al. Tyrosine phosphorylation of mitochondrial pyruvate dehydrogenase kinase 1 is important for cancer metabolism. Mol Cell 2011; 44: 864–877.
Morrison KC, Hergenrother PJ . Whole cell microtubule analysis by flow cytometry. Anal Biochem 2012; 420: 26–32.
Ng DC, Lin BH, Lim CP, Huang G, Zhang T, Poli V et al. Stat3 regulates microtubules by antagonizing the depolymerization activity of stathmin. J Cell Biol 2006; 172: 245–257.
Jin L, Li D, Alesi GN, Fan J, Kang HB, Lu Z et al. Glutamate dehydrogenase 1 signals through antioxidant glutathione peroxidase 1 to regulate redox homeostasis and tumor growth. Cancer Cell 2015; 27: 257–270.
We acknowledge Dr Anthea Hammond for editorial assistance. We thank the shared resources facilities of the Winship Cancer Institute and the Integrated Cellular Imaging Core of Emory University. We also acknowledge and thank Dr Shi-Yong Sun for his insightful scientific input in the development of this work. This work was supported in part by NIH grants R01 CA175316 (to SK), F31 CA183365 (to GNA) and ACS grant RSG-11-081-01 (to SK). GNA is an NIH predoctoral fellow. FRK and SK are Georgia Cancer Coalition Scholars. SK is a Robbins Scholar and an American Cancer Society Basic Research Scholar.
YK, ZGC, DMS and FRK provided critical reagents. LJ performed IHC staining and BLI study. KRM performed histopathological study. GNA and DL performed all the other experiments. GNA, LJ and SK designed the study and wrote the manuscript.
The authors declare no conflict of interest.
Supplementary Information accompanies this paper on the Oncogene website
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Alesi, G., Jin, L., Li, D. et al. RSK2 signals through stathmin to promote microtubule dynamics and tumor metastasis. Oncogene 35, 5412–5421 (2016). https://doi.org/10.1038/onc.2016.79
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